【摘要】 背景与目的:早期的研究显示,Notch1信号途径与肿瘤的发生密切相关,在细胞的生长、增殖、分化和凋亡中起着十分重要的作用。本研究旨在研究Notch1基因在食管鳞癌EC9706细胞中的表达及其对EC9706细胞凋亡的影响。方法:通过免疫细胞化学方法检测Notch1基因在EC9706细胞中的表达。采用RT-PCR技术扩增Notch1基因,构建真核表达载体pcDNA3.1-Notch1(命名为pcNICD),转染EC9706细胞,利用RT-PCR及Western blot检测稳定转染pcNICD载体、转染pcDNA3.1空载体及未处理的EC9706细胞中Notch1的表达,另通过流式细胞仪检测未处理、转染pcDNA3.1和转染pcNICD的EC9706细胞的凋亡。结果:EC9706细胞中发现Notch1基因的表达。与未处理的和转染pcDNA3.1的EC9706细胞相比,转染pcNICD的EC9706细胞中Notch1基因的mRNA和蛋白表达水平均明显增加,大约增加3倍(P<0.05);但未处理的和转染pcDNA3.1的EC9706细胞中Notch1的表达差异无统计学意义(P>0.05)。流式细胞检测结果显示,在未处理的和转染pcDNA3.1的EC9706细胞中,细胞凋亡率差异无统计学意义(P>0.05);但与与未处理的和转染pcDNA3.1的EC9706细胞相比,转染pcNICD的EC9706细胞转染后24h、48h和72h时细胞凋亡率明显增加(P<0.01)。结论:Notch信号途径的激活引起食管鳞癌EC9706细胞的凋亡,提示Notch1基因有可能成为治疗食管鳞癌的新靶点。更多还原

【Abstract】 BACKGROUND & OBJECTIVE: Notch1 signaling pathway is closely associated with carcinogenesis and plays a key role in cell growth, proliferation, differentiation, and apoptosis. This study was to investigate the expression of Notch1 gene in esophageal squamous cell carcinoma (ESCC) cell line EC9706 and its effects on cell apoptosis. METHODS: The expression of Notch1 gene in EC9706 cells was detected by immunocytochemistry. Notch1 gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR). A eukaryotic expression vector pcDNA3.1-Notch1 (termed pcNICD) was constructed and transfected into EC9706 cells; pcDNA3.1 was also transfected as control. After transfection, the expression of Notch1 in EC9706 cells was detected by RT-PCR and Western blot, cell apoptosis was measured by flow cytometry (FCM). RESULTS: The expression of Notch1 gene was detected in EC9706 cells. The mRNA and protein expression of Notch1 gene in pcNICD-transfected EC9706 cells were significantly increased by about 3 folds of those in untreated and pcDNA3.1-transfected cells (P<0.01). However, there was no difference in Notch1 expression between untreated and pcDNA3.1-transfected EC9706 cells (P>0.05). The apoptosis rate was significantly higher in pcNICD-transfected EC9706 cells than in untreated and pcDNA3.1-transfected cells (P<0.01); there was no prominent difference between untreated and pcDNA3.1-transfected EC9706 cells (P>0.05). CONCLUSION: The activated Notch1 signaling pathway gives rise to the apoptosis of EC9706 cells, suggesting that Notch1 gene may be a new therapeutic target for ESCC.更多还原